Protein denaturation, protein stability - Measuring protein unfolding - Circulating dichroismisn

6 important questions on Protein denaturation, protein stability - Measuring protein unfolding - Circulating dichroismisn

What kind of information can you obtain form Fluorescence Near-UV circular dichroism?

The amount of alfa-helix, beta-sheat and random coil in your sample and from this you can determine how much your sample is unfolded.

This is due to the different ways that these complexes absorbe light.

So the secondary structure

What is cirular dichroism?

A method to measure protein unfolding (CD).
Light travels in two directions or polarizations, namely: perpendicular (s-polarization) and parallel (p-polarization).

Expressed as the ellipticity.
The extent to which the polarization of the light is affected by the chiral atoms depends on the location of the atoms and the polarity of its environment.

What kind of information can you obtain from Near UV circular Dichroism?

This again looks at the Phenylalanin, Tyrosine and Tryptophan. And how much light is absorbed.
So the tertiary structure
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Which 2 methods are there for circular dichroism?

  • Secondary structure: Far-UV CD
  • Tertiary structure: Near-UV CD

Why is there a difference between protein unfolding between the 2 Circular Dichroism methods?

The tertystructure unfolds before the secondary structure. So that is why the fluoresence gives a unfolding befor the the Far.

Why is there a difference in protein unfolding between the 2 Circular Dichroism methods?

The tertiary structure unfolds before the secondary structure. So that is why the fluorescence gives an unfolding before the Far-UV CD.

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